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Targeting ULK1 and USP20 to modulate autophagy and chemosensitivity in cancer cell lines

  • Authors:
    • Tuqa Abu Thiab
    • Malek Zihlif
    • Dana Alqudah
    • Amer Imraish
  • View Affiliations / Copyright

    Affiliations: Department of Biological Sciences, School of Science, The University of Jordan, Amman 11942, Jordan, Department of Pharmacology, School of Medicine, The University of Jordan, Amman 11942, Jordan, Cell Therapy Center, The University of Jordan, Amman 11942, Jordan
    Copyright: © Abu Thiab et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 51
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    Published online on: February 25, 2026
       https://doi.org/10.3892/br.2026.2124
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Abstract

Autophagy is a conserved catabolic process essential for maintaining cellular homeostasis by degrading and recycling damaged organelles and misfolded proteins. In cancer, autophagy plays a dual role, acting as both a tumor suppressor and promoter depending on the stage and context. Unc‑51‑like kinase 1 (ULK1), a key initiator of autophagy, is tightly regulated by USP20, a de‑ubiquitinase that stabilizes ULK1 by preventing its lysosomal degradation. However, their roles in cancer progression and treatment response remain poorly understood. The present study investigated the baseline expression of ULK1 and USP20 across several cancer cell lines and evaluates the effects of their silencing on chemosensitivity. The findings of the present study showed that ULK1 was highly expressed in MCF‑7 breast cancer cells and minimally in U87 glioblastoma cells. USP20 showed high expression in MCF‑7, MDA‑MB‑231 and HepG2, and low expression in others. Combined silencing of ULK1 and USP20 with chemotherapy altered drug sensitivity across cancer types. ULK1 knockdown increased drug sensitivity and induced cell death in HepG2, MDA‑MB‑231 and PanC1 cell lines, but conferred chemoresistance in MCF‑7, A549 and U87 cancer cells. Similarly, USP20 silencing sensitized MCF‑7, HepG2 and PanC1 cells to chemotherapy, while enhancing survival in U87 cells. These results suggest that ULK1 and USP20 have cancer‑type‑specific roles in modulating autophagy and chemotherapy response. Targeting these proteins may provide novel therapeutic strategies to overcome chemoresistance and promote apoptosis in cancer treatment.
View Figures

Figure 1

Baseline expression of ULK1 and USP20
in different cancer cell line. Triplicates of normal untreated
cells were harvested, and the level of expression was determined
using the reverse transcription-quantitative PCR technique. (A)
Differential expression of ULK1 in each cancer cell line relative
to its expression in normal fibroblasts. (B) Differential
expression of USP20 in each cancer cell line relative to its
expression in normal fibroblasts. Data are presented as the mean ±
SEM. Statistical significance was indicated as
*P<0.05, **P<0.01 and
***P<0.001 (one-way ANOVA followed by Bonferroni's
multiple comparisons post hoc test). ULK1, Unc-51-like kinase
1.

Figure 2

Gene expression of ULK1 and USP20
genes after treatment with Lipofectamine coated siRNAs via reverse
transcription-quantitative PCR. (A) Fold change in the expression
of ULK1 after treatment with Lipofectamine-coated siULK1 at
different concentrations on cancer cells. (B) Fold change in the
expression of USP20 after the treatment with Lipofectamine-coated
siUSP20 at different concentrations on cancer cells. Statistical
significance was indicated as **P<0.01,
***P<0.001 and ****P<0.0001. ULK1,
Unc-51-like kinase 1; si-, small interfering.

Figure 3

Changes in the basal autophagy flux
in different cancer cell lines in response to doxorubicin. HepG2,
MCF-7, MDA-MB-231, U87 and normal fibroblast cells were treated
with different concentrations of doxorubicin chemotherapy. The
level of expression of ULK1, USP20, and P62 was determined
via reverse transcription-quantitative PCR. (A-C) Relative
expression of ULK1, USP20 and P62 in HepG2 cells. (D-F) Relative
expression of ULK1, USP20 and P62 in MCF-7 breast cancer cells.
(G-I) Relative expression of ULK1, USP20 and P62 in MDA-MB-231
breast cancer cells. (J-L) Relative expression of ULK1, USP20 and
P62 in U87 cells. (M-O) Relative expression of ULK1, USP20 and P62
in normal fibroblast cells. Three repeats were considered for each
treatment group. Data are presented as the mean ± SEM. Statistical
significance was indicated as *P<0.05,
**P<0.01, ***P<0.001 and
****P<0.0001 (one-way ANOVA followed by Bonferroni's
multiple comparisons post hoc test). ULK1, Unc-51-like kinase
1.

Figure 4

Changes in the basal autophagy flux
in A549 lung cancer cells and normal fibroblasts in response to
cisplatin chemotherapy treatment. Cells were treated with different
concentrations of cisplatin chemotherapy. The level of expression
of ULK1, USP20 and P62 was determined via reverse
transcription-quantitative PCR. (A-C) Relative expression of ULK1,
USP20, and P62 in A549 lung cancer cells. (D-F) Relative expression
of ULK1, USP20 and P62 in normal fibroblast cells. Three repeats
were considered for each treatment group. Data are presented as the
mean ± SEM. Statistical significance was indicated as
*P<0.05 and ***P<0.001 (one-way ANOVA
followed by Bonferroni's multiple comparisons post hoc test). ULK1,
Unc-51-like kinase 1.

Figure 5

Changes in the basal autophagy flux
in PanC1 pancreatic cancer cells and normal fibroblasts in response
to gemcitabine chemotherapy treatment. Cells were treated with
different concentrations of gemcitabine chemotherapy. The level of
expression of ULK1, USP20 and P62 was determined via reverse
transcription-quantitative PCR. (A-C) Relative expression of ULK1,
USP20 and P62 in PanC1 pancreatic cancer cells. (D-F) Relative
expression of ULK1, USP20 and P62 in normal fibroblast cells. Three
repeats were considered for each treatment group. Data are
presented as the mean ± SEM. Statistical significance was indicated
as *P<0.05, **P<0.01 and
***P<0.001 (one-way ANOVA followed by Bonferroni's
multiple comparisons post hoc test). ULK1, Unc-51-like kinase
1.
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Copy and paste a formatted citation
Spandidos Publications style
Abu Thiab T, Zihlif M, Alqudah D and Imraish A: Targeting ULK1 and USP20 to modulate autophagy and chemosensitivity in cancer cell lines. Biomed Rep 24: 51, 2026.
APA
Abu Thiab, T., Zihlif, M., Alqudah, D., & Imraish, A. (2026). Targeting ULK1 and USP20 to modulate autophagy and chemosensitivity in cancer cell lines. Biomedical Reports, 24, 51. https://doi.org/10.3892/br.2026.2124
MLA
Abu Thiab, T., Zihlif, M., Alqudah, D., Imraish, A."Targeting ULK1 and USP20 to modulate autophagy and chemosensitivity in cancer cell lines". Biomedical Reports 24.4 (2026): 51.
Chicago
Abu Thiab, T., Zihlif, M., Alqudah, D., Imraish, A."Targeting ULK1 and USP20 to modulate autophagy and chemosensitivity in cancer cell lines". Biomedical Reports 24, no. 4 (2026): 51. https://doi.org/10.3892/br.2026.2124
Copy and paste a formatted citation
x
Spandidos Publications style
Abu Thiab T, Zihlif M, Alqudah D and Imraish A: Targeting ULK1 and USP20 to modulate autophagy and chemosensitivity in cancer cell lines. Biomed Rep 24: 51, 2026.
APA
Abu Thiab, T., Zihlif, M., Alqudah, D., & Imraish, A. (2026). Targeting ULK1 and USP20 to modulate autophagy and chemosensitivity in cancer cell lines. Biomedical Reports, 24, 51. https://doi.org/10.3892/br.2026.2124
MLA
Abu Thiab, T., Zihlif, M., Alqudah, D., Imraish, A."Targeting ULK1 and USP20 to modulate autophagy and chemosensitivity in cancer cell lines". Biomedical Reports 24.4 (2026): 51.
Chicago
Abu Thiab, T., Zihlif, M., Alqudah, D., Imraish, A."Targeting ULK1 and USP20 to modulate autophagy and chemosensitivity in cancer cell lines". Biomedical Reports 24, no. 4 (2026): 51. https://doi.org/10.3892/br.2026.2124
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